Dying light: silent is differentiation tonight

UM > Faculty of Health Sciences > DEPARTMENT OF BIOMEDICAL SCIENCES

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	Dying light: silent is differentiation tonight
	ZHANG WENQI; XIE RUIYU
	2024-06-15
Size of Audience	250
Type of Speaker	poster presentation
Abstract	Inducible gene expression systems are useful tools across diverse research areas, including functional genomics, gene therapy, and tissue engineering. The commonly used Tet-On gene expression system, composed of a tetracycline-responsive element (TRE) promoter, a reverse tetracycline-controlled responsive transcriptional activator (rtTA), and a tetracycline-controlled transcriptional silencer (tTS), enables low background induction and precise control of gene expression. Upon exposure to tetracycline or its analogs such as doxycycline (dox), tTS undergoes a conformational change, liberating the TRE promoter for rtTA binding and initiating target gene expression. In this study, we employed an all-in-one Tet-On inducible PiggyBac (PB) transposon vector to trigger the expression of mKate2 red fluorescent protein upon dox administration during the differentiation of human embryonic stem cells (hESCs). Surprisingly, we observed a complete cessation of transgene induction during hESC differentiation into cardiomyocytes, pancreatic endoderm, and PAX6+ neuroectoderm lineages. This loss of transgene induction emerged early in the differentiation process, exhibiting lineage-specific differences among endoderm, mesoderm, and ectoderm derivatives. We further elucidated the significant role of tTS in exacerbating transgene silencing during differentiation. Intriguingly, unlike hESC differentiation, the inducibility of transgenes remained largely unaffected during the differentiation of mouse fibroblasts into adipocytes, indicating complex mechanisms underlying transgene silencing during hESC differentiation. While sporadic instances of transgene silencing during hESC differentiation have been reported, the near-complete loss of gene induction attributed to tTS during this process has remained unexplored until now. Our present study, which thoroughly examines this phenomenon and comprehensively evaluates inducible vectors across diverse differentiation conditions, offers crucial insights into the fields of mammalian cell engineering, gene therapy, and basic biology research.
Conference Date	2024-06-15
Conference Place	The 10th Macau Symposium on Biomedical Sciences, Macau
Document Type	Presentation
Collection	DEPARTMENT OF BIOMEDICAL SCIENCES
Corresponding Author	XIE RUIYU
Affiliation	Faculty of Health Sciences, University of Macau, Macau SAR, China
Recommended Citation GB/T 7714	ZHANG WENQI,XIE RUIYU. Dying light: silent is differentiation tonight, 2024-06-15.

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